Preparation and characterization of highly radioactive in vitro labeled adenovirus DNA and DNA restriction fragments

Biochemistry ◽  
1977 ◽  
Vol 16 (20) ◽  
pp. 4478-4483 ◽  
Author(s):  
Jesse K. Mackey ◽  
Karl H. Brackmann ◽  
Michael R. Green ◽  
Maurice Green
Keyword(s):  
Restriction Fragments ◽  
Dna Restriction Fragments ◽  
Dna Restriction

scholarly journals Effect of reversed orientation and length of An.Tn DNA bending sequences in the -35 and spacer domains of a consensus-like Escherichia coli promoter on its strength in vivo and gross structure of the open complex in vitro.

1996 ◽  
Vol 43 (1) ◽  
pp. 265-279 ◽  
Author(s):  
T Loziński ◽  
K L Wierzchowski
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Promoter Strength ◽  
Restriction Fragments ◽  
E Coli ◽  
Free Dna ◽  
Dna Repeat ◽  
Dna Restriction Fragments ◽  
Dna Restriction

In continuation of an earlier study (Loziński et al., 1991 Nucleic Acids Res. 19, 2947-2953) a series of consensus-like E. coli promoters with bending An.Tn sequences of different length (n = 3-8) and orientation in the -35 and spacer domains was constructed, cloned into the plasmid pDS3 and their strength in vivo measured in relation to an internal transcriptional standard. Gel mobilities of free DNA restriction fragments carrying these promoters and of open transcriptional complexes with cognate RNA polymerase were determined by polyacrylamide gel electrophoresis and the gross structure of the complexes interpreted in terms of the theoretically predicted superstructure of DNA restriction fragments. The results obtained together with those reported earlier show that bending of the DNA helix axis immediately upstream of the -35 domain generally lowers the promoter strength in vivo and brings about shortening of the mean square end-to-end distance between free DNA ends in the open complex in vitro. T4(-34 ...-37) and T5(-34 ...-38) tracts located in the nontemplate DNA strand had the largest and comparable effect on the promoter strength, while the A5.T5(-37 ... -41) sequence in either orientation (A5 tract in the template or nontemplate strand) exerted a much smaller effect. Promoters with the spacer bent by about 40 degrees but in different directions, by two A(n) (n = 5 or 6) tracts aligned in phase with the B-DNA repeat and located either in the template or nontemplate strands, had somewhat lower strength in vivo but the gross geometry of the respective open complexes was the same as that of a control promoter with straight spacer. Implications of these findings are discussed in connection with the existing model of E. coli transcriptional open complex.

Differentiation of two mouse cell lines is associated with hypomethylation of their genomes

1984 ◽  
Vol 4 (9) ◽  
pp. 1800-1806
Author(s):  
T H Bestor ◽  
S B Hellewell ◽  
V M Ingram
Keyword(s):  
Dna Methyltransferase ◽  
Cell Types ◽  
Mouse Cell ◽  
F9 Cells ◽  
Dna Restriction Fragments ◽  
Dna Restriction ◽  
F9 Embryonal Carcinoma Cells ◽  
Murine Erythroleukemia ◽  
Kinetics Of

Methyl-accepting assays and a sensitive method for labeling specific CpG sites have been used to show that the DNA of F9 embryonal carcinoma cells decreases in 5-methylcytosine content by ca. 9% during retinoic acid-induced differentiation, whereas the DNA of dimethyl sulfoxide-induced Friend murine erythroleukemia (MEL) cells loses ca. 3.8% of its methyl groups. These values correspond to the demethylation of 2.2 X 10(6) and 0.9 X 10(6) 5'-CpG-3' sites per haploid genome in differentiating F9 and MEL cells, respectively. Fluorography of DNA restriction fragments methylated in vitro and displayed on agarose gels showed that demethylation occurred throughout the genome. In uninduced F9 cells, the sequence TCGA tended to be more heavily methylated than did the sequence CCGG, whereas this tendency was reversed in MEL cells. The kinetics of in vitro DNA methylation reactions catalyzed by MEL cell DNA methyltransferase showed that substantial numbers of hemimethylated sites accumulate in the DNA of terminally differentiating F9 and MEL cells, implying that a partial loss of DNA-methylating activity may accompany terminal differentiation in these two cell types.

scholarly journals LOCALIZATION IN THE TOMATO GENOME OF DNA RESTRICTION FRAGMENTS CONTAINING SEQUENCES HOMOLOGOUS TO THE rRNA (45S), THE MAJOR CHLOROPHYLL a/b BINDING POLYPEPTIDE AND THE RIBULOSE BISPHOSPHATE CARBOXYLASE GENES

Genetics ◽  
1986 ◽  
Vol 112 (1) ◽  
pp. 93-105
Author(s):  
C E Vallejos ◽  
S D Tanksley ◽  
R Bernatzky
Keyword(s):  
Chlorophyll A ◽  
Ribosomal Rna ◽  
Chromosome 8 ◽  
Ribulose Bisphosphate Carboxylase ◽  
Chromosome 2 ◽  
Ribulose Bisphosphate ◽  
Bisphosphate Carboxylase ◽  
Restriction Fragments ◽  
Dna Restriction Fragments ◽  
Dna Restriction

ABSTRACT DNA restriction fragments containing sequences homologous to the ribosomal RNA (45s), the major chlorophyll a/b binding polypeptide (CAB) and the small subunit of ribulose bisphosphate carboxylase (RBCS) genes have been localized and mapped in the tomato nuclear genome by linkage analysis. Ribosomal RNA genes map to a single locus, R45s, which resides in a terminal position on the short arm of chromosome 2 and corresponds to the Nucleolar Organizer Region. The size of the 45s repeating unit is estimated to be approximately 9 kb in Lycopersicon esculentum and 11 kb in Lycopersicon pennellii. Five loci were found to contain CAB sequences. Two of the loci, Cab-1 (chromosome 2) and Cab-3 (chromosome 8), together accounted for more than 80% of the hybridization signal. These loci contain more than one CAB structural gene. The other three loci, Cab-2 (chromosome 8), Cab-4 (chromosome 7) and Cab-5 (chromosome 12), each account for <10% of the total signal and may contain only a single copy of the CAB structural sequence. Three loci were found to contain RBCS sequences. Rbcs-2 (chromosome 3) and Rbcs-3 (chromosome 2) were responsible for >80% of the signal, with the remainder being associated with Rbcs-1 (chromosome 2). Rbcs-2 and Rbcs-3 may contain more than one copy of the gene.

A sizing artifact of DNA restriction fragments with unusually long single-stranded termini

Gene ◽  
1991 ◽  
Vol 102 (1) ◽  
pp. 79-81 ◽  
Author(s):  
David M. Woodcock ◽  
Penelope J. Crowther ◽  
Judith P. Doherty ◽  
Martha E. Linsenmeyer ◽  
Detlev H. Krüger
Keyword(s):  
Restriction Fragments ◽  
Dna Restriction Fragments ◽  
Dna Restriction

Separation of DNA restriction fragments by ion-pair chromatography

1986 ◽  
Vol 359 ◽  
pp. 265-274 ◽  
Author(s):  
Stig Eriksson ◽  
Gunnar Glad ◽  
Per-Åke Pernemalm ◽  
Elisabeth Westman
Keyword(s):  
Ion Pair ◽  
Restriction Fragments ◽  
Ion Pair Chromatography ◽  
Dna Restriction Fragments ◽  
Dna Restriction
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